mRIPA lysis buffer

mRIPA is a good, all-purpose lysis buffer for solubilization of proteins from eukaryotic cells and tissue. For total protein preparation from adherent cells, pipet mRIPA directly onto cells grown on tissue culture plates and collect cells by scraping. Incubate homogenates on ice for 5 minutes, then centrifuge at 13,000 rpm in a microcentrifuge to pellet particulate material (DNA, etc). The supernatant contains solubilized protein.

Final concentrations:

  • 50 mM Tris pH 7.4
  • 1% NP-40
  • 0.25% deoxycholate
  • 150 mM NaCl

For 500 ml:

  • 25 ml 1M Tris/HCl pH 7.4
  • 5 ml NP-40
  • 1.25 g sodium deoxycholate
  • 4.38 g NaCl
  • Add H2O to 450 ml and re-adjust pH to 7.4
  • H2O to 500 ml

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